NHDL, a recombinant VL/VH hybrid antibody control for IgG2/4 antibodies

Corinna Lau; Martin Berner McAdam; Grethe Bergseth; Algirdas Grevys; Jack Ansgar Bruun; Judith Krey Ludviksen; Hilde Fure; Terje Espevik; Anders Moen; Jan Terje Andersen; Tom Eirik Mollnes

doi:10.1080/19420862.2019.1686319

NHDL, a recombinant V_L/V_H hybrid antibody control for IgG2/4 antibodies

MAbs. 2020 Jan-Dec;12(1):1686319. doi: 10.1080/19420862.2019.1686319.

Authors

Corinna Lau¹, Martin Berner McAdam², Grethe Bergseth¹, Algirdas Grevys^{2

3}, Jack Ansgar Bruun⁴, Judith Krey Ludviksen¹, Hilde Fure¹, Terje Espevik⁵, Anders Moen⁶, Jan Terje Andersen^{2

7}, Tom Eirik Mollnes^{1

5

8

9}

Affiliations

¹ Research Laboratory, Nordland Hospital Trust, Bodø, Norway.
² Department of Immunology, Oslo University Hospital-Rikshospitalet, and Centre for Immune Regulation, Oslo, Norway.
³ Centre for Immune Regulation and Department of Biosciences, University of Oslo, Oslo, Norway.
⁴ Department of Medical Biology, Proteomics Platform, University of Tromsø, Tromsø, Norway.
⁵ Centre of Molecular Inflammation Research, and Department of Cancer Research and Molecular Medicine, Norwegian University of Science and Technology, Trondheim, Norway.
⁶ Department of Biosciences, Proteomics core facility, University of Oslo, Oslo, Norway.
⁷ Department of Pharmacology, Institute of Clinical Medicine, Oslo University Hospital and University of Oslo, Oslo, Norway.
⁸ Department of Immunology, Oslo University Hospital, University of Oslo, Oslo, Norway.
⁹ Faculty of Health Sciences and K. G. Jebsen TREC, University of Tromsø, Tromsø, Norway.

Abstract

The mechanism of action of recombinant IgG2/4 antibodies involves blocking of their target without the induction of effector functions. Examples are eculizumab (Soliris®), which is used clinically to block complement factor C5, as well as anti-human CD14 (r18D11) and anti-porcine CD14 (rMIL2) produced in our laboratory. So far, no proper IgG2/4 control antibody has been available for controlled validation of IgG2/4 antibody functions. Here, we describe the design of a recombinant control antibody (NHDL), which was generated by combining the variable light (V_L) and heavy (V_H) chains from two unrelated specificities. NHDL was readily expressed and purified as a stable IgG2/4 antibody, and showed no detectable specificity toward any putative antigen present in human or porcine blood. The approach of artificial V_L/V_H combination may be adopted for the design of other recombinant control antibodies.

Keywords: IgG2/4; Recombinant antibody; control antibody; hybrid; therapeutic antibody.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Antibodies, Monoclonal / genetics*
Antibodies, Monoclonal / metabolism
Biological Therapy
Blood Proteins / metabolism
Epitopes / metabolism
Humans
Immunoglobulin G / genetics*
Immunoglobulin Heavy Chains / genetics*
Immunoglobulin Light Chains / genetics*
Lipopolysaccharide Receptors / immunology
Lipopolysaccharide Receptors / metabolism
Mice
Placebos
Protein Engineering
Recombinant Fusion Proteins / genetics*
Swine

Substances

Antibodies, Monoclonal
Blood Proteins
Epitopes
Immunoglobulin G
Immunoglobulin Heavy Chains
Immunoglobulin Light Chains
Lipopolysaccharide Receptors
Placebos
Recombinant Fusion Proteins

Grants and funding

TThis work was supported by Helse Sør-Øst RHF [2018052]; [Norges Forskningsråd [287927]; Norges Forskningsråd [179573]; Norwegian council of cardiovascular disease [NCCD-2014]; Helse Nord RHF [8416/SFP1083-13]; Odd Fellow Foundation [OFF-2015].